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- Table of Contents
TPMT · Western blot design guide
TPMT Western Blot Planning Guide
Plan a TPMT Western blot around the catalog-observed 28.2 kDa band, image-backed A00671-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
TPMT Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | 28.2 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Parathyroid gland | |
| Negative control | Bone marrow |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 28.2 kDa |
| ⓘ | Localization | Cytoplasm |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated TPMT Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Kidney |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, short transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A00671-1 at datasheet starting dilution |
| Primary incubation | Overnight at 4 °C with gentle agitation |
| Secondary | Species-matched HRP conjugate at validated dilution |
| Wash | 3 × 5 min in TBST |
| Detection | Chemiluminescent substrate |
| Exposure | Bracket exposures to avoid saturation |
Section 2
What Is the Expected TPMT Western Blot Band Size?
Use the product-observed 28.2 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 28.2 kDa | Matches the authoritative product WB observation. |
| 28.2 kDa calculated | Use as UniProt context, not as a replacement observed band. |
| Unexpected additional signal | Do not assign identity without orthogonal positive/negative controls. |
💡Expected TPMT appearancePlan around 28.2 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 28.2 kDa; use this as the primary experimental expectation. |
| Calculated mass | 28.2 kDa from UniProt P51580; retain as context. |
| Gel selection | 12-15%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A00671-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 28.2 kDa | Matches the authoritative product WB observation. | Confirm with orthogonal controls and the linked product record. |
| Additional band | May reflect processing, modification, or non-specific signal. | Run a dilution series and compare positive/negative controls. |
| Weak signal | Target abundance or transfer may be limiting. | Verify transfer, increase positive-control abundance, and bracket exposure. |
Experiment evidence
Western blot images showing TPMT band
A00671-1 · TPMT
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TPMT Western blot
🧪Use Parathyroid gland as the first positive-control candidate and Bone marrow as the HPA Not detected negative candidate.
Positive control: Parathyroid gland (High)
Negative control: Bone marrow (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for TPMT
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Parathyroid gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Thyroid gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Kidney | Reported tissue cells | High | Protein (HPA) | HPA → |
| Skin | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced TPMT Western Blot Tips
Deeper troubleshooting and optimisation questions for TPMT, answered from its protein features.
Which band should guide the blot?
Use 28.2 kDa, the observation attached to the authoritative A00671-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 28.2 kDa expectation.
Which positive control should I start with?
Start with Parathyroid gland, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Bone marrow as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for TPMT Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00671-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
Compare only the supplied target-verified records: PMC11679344, PMC2815203, PMC2583164.
Boster reagents
TPMT Western Blot Reagents
Human/Mouse/Rat-reactive TPMT Western blot reagents with authoritative product imagery.
Real WB data
Anti-TPMT Antibody Picoband®
Cat # A00671-1
Only image-backed, WB-validated Human/Mouse/Rat recommendations from the prepared catalog evidence are shown.
Source: prepared picoband-wb product evidence; each card retains its own SKU, URL, observed band, and authoritative WB image.
References
- UniProt P51580
- Human Protein Atlas — TPMT
- A00671-1 product record
- PMC11679344 — A Rapid and Reliable Absorbance Assay to Identify Drug-Drug Interactions with Thiopurine Drugs.
- PMC2815203 — Thiopurine S-methyltransferase pharmacogenetics: functional characterization of a novel rapidly degraded variant allozyme.
- PMC2583164 — Thiopurine S-methyltransferase pharmacogenetics: autophagy as a mechanism for variant allozyme degradation.